Short/small hairpin RNA molecules

An RNA polymerase, hairpin technology, applied in the field of molecular biology, can solve environmental problems, insufficient insect control of insecticides, etc.

Pending Publication Date: 2021-01-08
DOW AGROSCIENCES LLC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Large insect larval populations, heavy rainfall, and inappropriate application of one or more insecticides can all lead to inadequate insect control
Furthermore, the sequentia

Method used

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  • Short/small hairpin RNA molecules
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  • Short/small hairpin RNA molecules

Examples

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example 1

[0305] Example 1: shRNA molecules contain novel structural features

[0306] Design and development of novel short / small hairpin RNA (shRNA) molecules to inhibit RNA transcribed from expressed genes within living cells of an organism. In some embodiments, shRNA molecules are single-stranded polynucleotide molecules that are less than 70 nucleotides (ie, <70 nt) in length. In other embodiments, shRNA molecules are single-stranded polynucleotide molecules that are less than 80 nucleotides (ie, <80nt) in length. In additional embodiments, shRNA molecules are single-stranded polynucleotide molecules that are less than 90 nucleotides (ie, <90nt) in length. Single-stranded shRNA molecules form a secondary structure called a stem-loop structure (eg, a hairpin loop structure). The shRNA stem-loop structure results from intramolecular nucleotide base pairing, such that a single-stranded shRNA molecule folds and forms base pairs with another part of the same shRNA polynucleotide stran...

example 2

[0308] Example 2: shRNA molecules contain hairpin structures

[0309] The hairpin structure of an shRNA is a stretch of primarily unpaired or single-stranded nucleotides of the shRNA polynucleotide that results when the shRNA polynucleotide folds and forms base pairs with another portion of the same strand of the shRNA polynucleotide. Although the hairpin remains primarily unpaired or single-stranded, some double-stranded junctions can form between nucleotides within the hairpin of the shRNA. It is not impossible to observe double-stranded fragments of 1, 2, 3, 4, or 5 nucleotides in length within the hairpin structure of the shRNA molecule. In addition, multiple double-stranded fragments can be observed within the hairpin structure of shRNA molecules. In some aspects, the hairpin structure of the shRNA molecule contains unstructured polynucleotide sequences. These unstructured polynucleotide sequences within the loop structure are single-stranded bulges / mismatches of single...

example 3

[0311] Example 3: shRNA molecules contain a stem structure

[0312] The stem structure of an shRNA is the double-stranded polynucleotide sequence of a single-stranded shRNA polynucleotide that results when the shRNA polynucleotide folds and forms a double-stranded base pair with another portion of the same strand of the shRNA polynucleotide. The stem structure remains predominantly paired or double-stranded, where no single-stranded polynucleotide sequence is formed between nucleotides within the stem structure of the shRNA due to unbonding of nucleotides. In some aspects, the stem structure of an shRNA molecule can contain unstructured polynucleotide sequences. In other aspects, the stem structure of the shRNA molecule can all assume a paired double-stranded structure. In additional aspects, shRNA stem structures can be designed to be greater than 16 nucleotides in length (ie >16 bp). In other aspects, shRNA stem structures can be designed to be less than or equal to 25 nuc...

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Abstract

This disclosure concerns compositions and methods for novel RNA inhibition molecules (e.g., small/short hairpin RNA (shRNA) molecules), polynucleotides encoding such molecules, use of such novel RNA inhibition molecules to inhibit a target gene by suppressing the expression of the mRNA of the target gene, and for application such as the of control insect pests, and transgenic plants that produce,and are protected, by these novel RNA inhibition molecules are described.

Description

[0001] This application claims priority to US 62 / 773,355, filed November 30, 2018, and US 62 / 633,720, filed February 22, 2018, both of which are hereby incorporated by reference in their entirety. [0002] Incorporation by reference of electronically submitted material [0003] Incorporated by reference in its entirety is the computer-readable nucleotide / amino acid sequence listing, filed at the same time and identified as follows: A 90.2 KB ASCII (text )document. technical field [0004] The present invention relates generally to the field of molecular biology as applied to agricultural science. More particularly, certain embodiments relate to compositions and methods for inhibiting a target gene by inhibiting the expression of its mRNA using short / small hairpin RNA (shRNA) molecules. In some embodiments, shRNA molecules are used for insect control. Also disclosed are methods of making and using shRNA molecules in the development of novel insecticidal molecules in transgen...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/82
CPCC12N15/113C12N15/8286C12N2310/14C12N2310/531G01N33/5085Y02A40/146C12N15/8218C12N2310/141C12N2310/533A01N63/60
Inventor E·菲什里维奇K·奈娃杨效曾M·L·弗雷M·兰加斯瓦米W·罗P·甘道夫P·M·沃特豪斯
Owner DOW AGROSCIENCES LLC
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